CN101732763A - Method for preparing colloidal/pasty allogeneic demineralized bone compound and application - Google Patents
Method for preparing colloidal/pasty allogeneic demineralized bone compound and application Download PDFInfo
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- CN101732763A CN101732763A CN200810172475A CN200810172475A CN101732763A CN 101732763 A CN101732763 A CN 101732763A CN 200810172475 A CN200810172475 A CN 200810172475A CN 200810172475 A CN200810172475 A CN 200810172475A CN 101732763 A CN101732763 A CN 101732763A
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Abstract
The invention relates to a method for preparing colloidal/creamy allogeneic demineralized bone compound with fluidity and application. As the product has the characteristics of certain fluidity, viscidity and strong plasticity, the product can be widely used as a filling material for treating bone defects caused by various causes clinically, particularly applicable to the filling of irregular-shape bone defects; the product is prepared by the following steps: using allogeneic bone of human being as a raw material, processing the allogeneic bone into bone grains with granularity of between 0.1 and 1.2 cm; and performing demineralization and freezing process on the bone grains to prepare the demineralized and frozen bone grains, mixing the prepared demineralized and frozen bone grains and biocompatible fluidic organism serving as a carrier into the fluidic colloidal or pasty allogeneic demineralized bone compound, or adding a thixotropic additive into the demineralized bone compound to make the demineralized bone compound more viscous and massive.
Description
Technical field
The present invention relates to a kind of preparation method and application that is used for the embedded material of the damaged filling of surgical intervention bone.Be specifically related to a kind of glue of flowable, paste, or the preparation method and the application of pasty state allogeneic demineralized bone complex.
Background technology
Widespread usage human homogeneous variant bone implant material substitutes from the body bone as the damaged bone grafting material that carries out repair and reconstruction of treatment bone both at home and abroad at present.Because this material has nature bone conductivity and osteoinductive, it is particularly suitable for the damaged filling of clinical general bone, the immediate union that promotes bone does not connect, spinal column or joint fusion, the filling that the oromaxillo-facial region bone is damaged, and is used to rebuild damaged filling bone grafting and the structural remodeling of osteoarthrosis that causes because of diseases such as tumor, wound, infection.Demineralized bone is osseous tissue to be carried out making after acid demineralization is handled calcium content reduces in a large number in the osseous tissue, thereby improves its osteoinductive, reaches the better clinical effectiveness that promotes the osteocyte growth.Demineralized bone is extensive use of at present abroad.Fill and dental applications and also begin to adopt demineralized bone bone meal to be used for small-sized bone cavity on the domestic market gradually, as the demineralization bone meal of Aorui Biological Material Co., Ltd., Shanxi's production.State known the office on February 16th, 2005 Granted publication a kind of " bone bone meal of the same race and preparation method thereof " (publication number is CN 1579561A) patent of invention, this Patent publish the preparation method of basic demineralization bone meal.Yet because most of bones are damaged clinically is shape lack of standardization, directly is filled with certain degree of difficulty and can not fully fills up damaged with dry demineralization bone meal or skeletal grain.Sometimes adopting patient's self-blood or normal saline to be mixed into the semi-fluid shape with dry bone meal or skeletal grain in the operation is convenient to fill.Because blood need now be adopted in operation and can not use on different patients without blood group coupling, also have the possibility of cross infection simultaneously, not ideal method.In view of above-mentioned the deficiencies in the prior art, the technical problem to be solved in the present invention is to make semi-fluid shape demineralized bone mixture can be directly used in clinical under the condition that does not change the demineralized bone biological property.The present domestic glue of not seeing flowable as yet, paste, or pasty state allogeneic demineralized bone complex is as the filling material of bone aspect for information about.
Summary of the invention
The objective of the invention is to disclose a kind of flowing, paste, or the preparation method and the application of pasty state allogeneic demineralized bone complex.The glue of this flowable, paste, or pasty state allogeneic demineralized bone complex good biocompatibility have nature bone conductivity and osteoinductive, and immunological rejection is little.Its main feature is that plasticity is strong, and the bone that can be used for filling various shapes lack of standardization is damaged.Need in operation process, not get blood or now mix with normal saline, easy to use, save operating time, reach ideal filling effect simultaneously, for better filling material of bone is provided clinically.
The glue of said this flowable, paste, or pasty state allogeneic demineralized bone complex make by the following method:
1) personnel selection allogeneic osseous tissue is as raw material.The preparation particle diameter is the skeletal grain of 0.1cm to 1.2cm;
Skeletal grain is carried out degreasing sterilization with ultrasonic method in ethanol and/or TritonX (TritonX-100) solution; Skeletal grain behind the degreasing sterilization or bone fibres are carried out the demineralization processing with ultrasonic method in hydrochloric acid or peracetic acid, the residue calcium content of the osseous tissue after demineralization is handled should be less than 2%.Use aseptic water washing, disacidify is raised to pH value between 6.5 to 7.5;
2) skeletal grain after the demineralization being carried out in a vacuum lyophilization handles;
3) the demineralization lyophilizing skeletal grain for preparing is mixed into the glue of flowable, paste, or pasty state allogeneic demineralized bone complex with the stream shape Organic substance with biocompatibility as carrier;
4), can in the demineralized bone complex, add again and have thixotropic additive and make the demineralized bone complex more sticky and thick and heavy in order to promote skeletal grain with its Organic substance carrier better mixing.
5) also can add other biochemistry composition in this demineralized bone complex according to different clinical needs as collagen protein, antibiotics, antitoxin, hormone, enzyme, bone morphogenetic protein(BMP) (BMPs), living cells such as human bone marrow cell and stem cell.
6) this demineralized bone complex can be pre-mixed under aseptic condition and preserve.Also the demineralization lyophilizing skeletal grain for preparing separately can be preserved with Organic substance carrier or other additive, remix uses in operation.
Preferred version of the present invention is: the ideal dimensions in skeletal grain footpath is that 0.2cm is to 1cm.During degreasing sterilization used concentration of alcohol be 70% or TritonX (TritonX-100) concentration be 0.5%, the used salt acid concentration is 0.6mol/L during demineralization, lyophilization pressure in-20 ℃ to-35 ℃ vacuum is to carry out under the 150-100mTorr condition.The desirable Organic substance carrier of complex is polymer and the esters derivative such as the list of the aqueous or solid many hydroxyls of shape (base), dibasic acid esters glycerol, propylene glycol, Polyethylene Glycol, acetin etc.Additive uses polyvinyl alcohol, gel, and gelatin, pectin, collagen protein, starch, methylcellulose etc. have thixotropic material makes mixture more sticky and thick and heavy.
Because the glue of this flowable, paste, or pasty state demineralized bone complex is through abundant defat, sterilization, and demineralization prepares after the lyophilization, has both effectively reduced immunogenicity, has kept osteogenic factor, has strengthened its osteoinductive energy again.The glue that adds the flowable that forms behind organic carrier and the additive, paste, or pasty state demineralized bone complex becomes the damaged desirable filling material of bone of clinical bone lack of standardization.It is damaged to be used for filling any capable shape bone, the cavity, and wound, the growth of inducing self-body osteocyte promotes the healing early of bone.
The specific embodiment
Below by embodiment the present invention is described in more detail, but content of the present invention not only is confined to the following examples.
1. available human cortical bone is processed in the preparation of skeletal grain, spongy bone, and skin spongy bone.
1.1 the processing of skeletal grain is to adopt present existing process technology, it is the granule of 0.2cm to 1cm that the footpath is made in human bone pulverizing.
2. degreasing sterilization is handled
Getting the skeletal grain that has made, to put into concentration be that 70% alcoholic solution shook defat 4 hours at normal temperatures.Bone material with the ratio of alcoholic solution is: every gram bone material needs the 30ML alcoholic solution.
3. demineralization is handled
3.1 the bone material behind the degreasing sterilization added in the hydrochloric acid solution that concentration is 0.6N shook demineralization at normal temperatures 3 hours.Bone material with the ratio of hydrochloric acid solution is: every gram bone material needs the 50ML hydrochloric acid solution.Filter out hydrochloric acid solution, add sterilized water again and wash repeatedly 3 to 5 times, filter is done.
Be immersed in the 0.1M buffer solution of sodium phosphate and be raised between 6.5 to 7.5 up to pH value 3.2 will filter bone material after doing, filter is done.Adding sterilized water again washes 3 to 5 times repeatedly.The residue calcium content of the osseous tissue after demineralization is handled should be less than 2%.
4. lyophilization is handled
It ℃ is to carry out under the 150-100mTorr condition lyophilization 24-48 hour to-35 ℃ of vacuum pressure in temperature-20 that bone material after demineralization handled is put into freezer dryer.
5. the preparation of glue/paste demineralized bone complex
5.1 the glycerol that the demineralized bone material 10g after the lyophilization and 50g aseptic injection are used fully mixes, and slowly stirs the mixture 1 hour or the longer time fully is mixed into paste up to complex.
5.2 the paste demineralized bone complex for preparing is preserved under aseptic condition.Also can be kept at becomes injection type in the asepsis injector.
About the embodiment several points to note:
1. the bone material after demineralization is handled also can be mixed with agglutination/paste demineralized bone complex without the glycerol that lyophilization is directly used with aseptic injection.
2. in mixed process, can at any time add other biochemistry composition by clinical needs.
Claims (5)
1. the preparation method and the application of glue/pasty allogeneic demineralized bone complex is characterized in that its preparation method is as follows:
(1) with human homogeneous allograph bone as raw material, being processed into particle diameter is the skeletal grain of 0.1cm to 1.2cm;
(2) the demineralized bone bone meal technology of preparing (state know innings publication number be CN1579561A) of employing to have carries out skeletal grain to carry out degreasing sterilization with ultrasonic method in the degreasing sterilization with ultrasonic method in ethanol and/or TritonX (TritonX-100) solution; Carrying out demineralization with ultrasonic method in hydrochloric acid or peracetic acid handles; Use aseptic water washing, disacidify is raised to pH value between 6.5 to 7.5;
(3) skeletal grain after the demineralization being carried out in a vacuum lyophilization handles.
(4) the demineralization lyophilizing skeletal grain for preparing is mixed into the glue of flowable, paste, or pasty state allogeneic demineralized bone complex with the stream shape Organic substance with biocompatibility as carrier;
(5), can in the demineralized bone complex, add again and have thixotropic additive and make the demineralized bone complex more sticky and thick and heavy in order to promote skeletal grain with its Organic substance carrier better mixing.
(6) also can add in this demineralized bone complex according to different clinical needs other biochemistry composition as:
Collagen protein, antibiotics, antitoxin, hormone, enzyme, bone morphogenetic protein(BMP) (BMPs), living cells such as human bone marrow cell and stem cell.
(7) this demineralized bone complex can be pre-mixed under aseptic condition and preserve.Also the demineralization lyophilizing skeletal grain for preparing separately can be preserved with Organic substance carrier or other additive, remix uses in operation.
2. glue as claimed in claim 1/pasty allogeneic demineralized bone complex is characterized in that: in the step (3)
Temperature is-20 ℃ to-35 ℃ when carrying out the lyophilization processing in the air, and pressure is 150-100mTorr in the vacuum, is 24 to 48 hours according to different bone material quantity cooling times.
3. glue as claimed in claim 1/pasty allogeneic demineralized bone complex, it is characterized in that: the stream shape Organic substance that biocompatibility is arranged together in the step (4) is polymer and the esters derivative such as the lists of the aqueous or solid many hydroxyls of shape (base), dibasic acid esters glycerol, propanol, Polyethylene Glycol, acetin etc.
4. glue as claimed in claim 1/pasty allogeneic demineralized bone complex is characterized in that: the thixotropic additive of tool is a polyvinyl alcohol in the step (5), gel, gelatin, pectin, collagen protein, starch, methylcellulose etc.
5. glue as claimed in claim 1/pasty allogeneic demineralized bone complex is characterized in that: the demineralized bone complex that step (7) gets togather in advance also can be kept at becomes injection type in the asepsis injector.
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| Application Number | Priority Date | Filing Date | Title |
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| CN200810172475A CN101732763A (en) | 2008-11-12 | 2008-11-12 | Method for preparing colloidal/pasty allogeneic demineralized bone compound and application |
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| CN200810172475A CN101732763A (en) | 2008-11-12 | 2008-11-12 | Method for preparing colloidal/pasty allogeneic demineralized bone compound and application |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102895703A (en) * | 2012-11-08 | 2013-01-30 | 北京鑫康辰医学科技发展有限公司 | Decalcified bone matrix material and preparation method thereof |
| CN106413767A (en) * | 2014-04-25 | 2017-02-15 | 生物银行公司 | Method for producing a bone paste |
| CN110075356A (en) * | 2019-03-28 | 2019-08-02 | 中国辐射防护研究院 | A kind of plasticity packing material and preparation method thereof for surgical intervention bone defect |
| CN111012947A (en) * | 2019-12-30 | 2020-04-17 | 南京财经大学 | Injectable and self-healing starch-based hydrogel and preparation method and application thereof |
| CN113015546A (en) * | 2018-08-29 | 2021-06-22 | 华沙整形外科股份有限公司 | Bone material hydration device and method |
| CN115645617A (en) * | 2022-09-09 | 2023-01-31 | 北京鑫康辰医学科技发展有限公司 | Preparation method of bone matrix gel with biological activity |
-
2008
- 2008-11-12 CN CN200810172475A patent/CN101732763A/en active Pending
Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102895703A (en) * | 2012-11-08 | 2013-01-30 | 北京鑫康辰医学科技发展有限公司 | Decalcified bone matrix material and preparation method thereof |
| CN106413767A (en) * | 2014-04-25 | 2017-02-15 | 生物银行公司 | Method for producing a bone paste |
| CN106413767B (en) * | 2014-04-25 | 2019-06-04 | 生物银行公司 | The method for producing bone mud |
| CN113015546A (en) * | 2018-08-29 | 2021-06-22 | 华沙整形外科股份有限公司 | Bone material hydration device and method |
| CN110075356A (en) * | 2019-03-28 | 2019-08-02 | 中国辐射防护研究院 | A kind of plasticity packing material and preparation method thereof for surgical intervention bone defect |
| CN110075356B (en) * | 2019-03-28 | 2021-08-17 | 中国辐射防护研究院 | Plastic filling material for surgical treatment of bone defects and preparation method thereof |
| CN111012947A (en) * | 2019-12-30 | 2020-04-17 | 南京财经大学 | Injectable and self-healing starch-based hydrogel and preparation method and application thereof |
| CN111012947B (en) * | 2019-12-30 | 2021-11-12 | 南京财经大学 | Injectable and self-healing starch-based hydrogel and preparation method and application thereof |
| CN115645617A (en) * | 2022-09-09 | 2023-01-31 | 北京鑫康辰医学科技发展有限公司 | Preparation method of bone matrix gel with biological activity |
| CN115645617B (en) * | 2022-09-09 | 2023-07-25 | 北京鑫康辰医学科技发展有限公司 | Preparation method of bone matrix gel with bioactivity |
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Application publication date: 20100616 |